Bioscience, Biotechnology, and Biochemistry
Online ISSN : 1347-6947
Print ISSN : 0916-8451
Biochemistry & Molecular Biology Preliminary Communication
Expression of a Functional Single-chain Antibody against GA24/19 in Transgenic Tobacco
Naoki SHIMADAYoshihito SUZUKIMasatoshi NAKAJIMAUdo CONRADNoboru MUROFUSHIIsomaro YAMAGUCHI
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1999 Volume 63 Issue 4 Pages 779-783

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Abstract
  An anti-gibberellin A24/19 single-chain Fv gene was constructed from γ and κ genes cloned from a hybridoma cell line producing monoclonal antibody against gibberellin A24/19, biosynthetic precursors of gibberellin A4/1 which are biologically active per se. The single-chain Fv gene was introduced into tobacco plants after the binding activity of the single-chain Fv expressed in Escherichia coli was confirmed. When the single-chain Fv expression is targeted to endoplasmic reticulum, the plants could accumulate the single-chain Fv protein with the antigen binding activity up to 3.6% of the total soluble protein. On the other hand, when the expression is targeted to cytosol, accumulation of the single-chain Fv protein was not detected at all. The dwarf phenotype of the transgenic plants expressing the single-chain Fv protein, together with the preliminary analytical data indicating a decreased level of gibberellin A1 in the dwarf transgenics, suggested that the single-chain Fv decreased the concentration of bioactive gibberellins by trapping and inhibiting the metabolism of gibberellin A24 and/or A19 to gibberellin A4 and/or A1.
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© 1999 by Japan Society for Bioscience, Biotechnology, and Agrochemistry
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